公开(公告)号：US20230349912A1

公开(公告)日：2023-11-02

申请号：US18026678

申请日：2021-09-17

Applicant: AMGEN INC.

Inventor： Gang XIAO ,  Pavel BONDARENKO ,  Liuqing SHI ,  Thomas M. DILLON ,  Da REN ,  Margaret Speed RICCI ,  Weidong CUI ,  John Joseph HARRAHY ,  Jake PAWLOWSKI ,  Andrew DYKSTRA ,  Dylan RIGGS

IPC: G01N33/68

CPC classification number: G01N33/6821 ,  G01N33/6848 ,  G01N2333/976

Abstract: Provided herein are methods of processing a polypeptide or protein for analysis, e.g., peptide mapping analysis by mass spectrometry. In exemplary embodiments, the method comprises incubating a digested sample at a mildly acidic pH and/or in the presence of a chaotrope, wherein the digested sample is produced by digesting a polypeptide with a protease to produce a digested sample comprising at least two peptides. In exemplary embodiments, the method comprises digesting the polypeptide with a protease which cleaves C-terminal to tryptophan to produce a digested sample comprising at least two peptides. In exemplary embodiments, the method comprises digesting the polypeptide with trypsin at an enzyme:substrate (E:S) weight ratio of about 1:1 to about 1:15 to produce a digested sample comprising at least two peptides. In exemplary aspects, the digested sample comprises at least one or two peptides each comprising a tyrosine at the C-terminus.

公开(公告)号：US20230235024A1

公开(公告)日：2023-07-27

申请号：US17865364

申请日：2022-07-14

Applicant: Amgen Inc.

Inventor： Zhimei DU ,  Pranhitha REDDY ,  Randal B. BASS ,  Feng HE ,  William C. FANSLOW, III ,  Yuling ZHANG ,  Da REN ,  Randall R. KETCHEM

IPC: C07K16/00

CPC classification number: C07K16/00 ,  C07K2317/41 ,  C07K2317/52 ,  C07K2317/524 ,  C07K2317/14 ,  C07K2319/30

Abstract: The present invention relates to variant Fc-containing molecules, such as antibodies and Fc-fusion molecules, having glycosylation characteristics favorable to large-scale production of therapeutic molecules containing such variant Fc.

公开(公告)号：US20210255075A1

公开(公告)日：2021-08-19

申请号：US16980344

申请日：2019-03-13

Applicant: Amgen Inc.

Inventor： Da REN ,  Melissa SATO

IPC: G01N1/40 ,  G01N33/68 ,  G01N30/06

Abstract: The disclosed methods are directed to preparing polypeptides for multi-attribute analysis. The polypeptides are optionally denatured, reduced, and/or alkylated before being subjected to a first digestion. Following the first digestion the large and small fragments resulting from the digestion are separated from each other. A second digestion is then performed on the larger of the fragments. All of the fragments from the two digestions are then analyzed chromatographically, electrophoretically, or spectrometrically, or a combination of these methods. The methods are especially useful for the preparation of therapeutic polypeptides for analysis, especially those that are not easily cleaved.

公开(公告)号：US20160207980A1

公开(公告)日：2016-07-21

申请号：US14916993

申请日：2014-09-05

Applicant: AMGEN INC.

Inventor： Zhimei DU ,  Pranhitha REDDY ,  Randal B. BASS ,  Feng HE ,  William C. FANSLOW, III ,  Yuling ZHANG ,  Da REN ,  Randal R. KETCHEM

IPC: C07K16/00

CPC classification number: C07K16/00 ,  C07K2317/14 ,  C07K2317/41 ,  C07K2317/52 ,  C07K2317/524 ,  C07K2319/30

Abstract: The present invention relates to variant Fc-containing molecules, such as antibodies and Fc-fusion molecules, having glycosylation characteristics favorable to largescale production of therapeutic molecules containing such variant Fc. Described herein are compositions and methods to improve glycosylation maturation of and to minimize the culture process-dependent effects of Fc-containing molecules, e.g., Fc-fusion molecules and antibodies. Creating single and multiple amino acid substitutions within the Fc domain with the aim to improve high mannose processing and glycosylation maturation.

Abstract translation: 本发明涉及具有有利于大规模生产含有这种变体Fc的治疗分子的糖基化特征的变体Fc-分子，例如抗体和Fc-融合分子。 本文描述的是改善糖基化成熟并使得含Fc分子例如Fc-融合分子和抗体的培养过程依赖性作用最小化的组合物和方法。 在Fc结构域内产生单个和多个氨基酸取代，目的是改善高甘露糖加工和糖基化成熟。

公开(公告)号：US20220404370A1

公开(公告)日：2022-12-22

申请号：US17834518

申请日：2022-06-07

Applicant: AMGEN INC.

Inventor： Da REN ,  Izydor APOSTOL ,  Alexander Julian VEACH

IPC: G01N33/68 ,  B01D15/34

Abstract: The disclosed methods are directed to detecting polypeptide fragments (“clips”) of parental polypeptides. Parental polypeptides and clips are optionally denatured and then fractionated using a matrix. After a first elution (high molecular weight fraction), an additional elution step retrieves a low molecular weight fraction containing clips. If the clips are an appropriate size for the targeted detection method, such as mass spectrometry, then analysis of this fraction proceeds separately from the high molecular weight fraction, or the clips fraction is mixed with the proteolyzed high molecular weight fraction before analysis. However, if the clips are too large for the intended analytical method(s), then the clips are also proteolyzed. The digested high molecular weight and low molecular weight fractions can be analyzed separately or combined. Analysis of combined samples favors clip quantitation because the clips are analyzed together with the remaining counterpart of the parental polypeptide.

公开(公告)号：US20190233543A1

公开(公告)日：2019-08-01

申请号：US16338292

申请日：2017-09-28

Applicant: AMGEN INC.

Inventor： Neeraj Jagdish AGRAWAL ,  Pavel BONDARENKO ,  Joon Hoi HUH ,  Andrew NICHOLS ,  Da REN ,  Richard SMITH ,  Riki STEVENSON

IPC: C07K16/40 ,  C07K16/28

CPC classification number: C07K16/40 ,  C07K16/2869 ,  C07K2317/21 ,  C07K2317/24 ,  C07K2317/52 ,  C07K2317/55 ,  C07K2317/90 ,  C07K2317/92 ,  C07K2317/94 ,  G01N11/14

Abstract: The present invention concerns a method for preparing antigen binding proteins with reduced viscosity. The method proceeds by replacing residues in high viscosity variable domain subfamilies with residues in correlating low viscosity subfamilies. The method further comprises substituting residues in the Fc domain with residues associated with low viscosity and adding charged residues to the C-terminus of the Fc domain. The present invention further concerns antigen binding proteins produced by this method.

公开(公告)号：US20190004060A1

公开(公告)日：2019-01-03

申请号：US16022850

申请日：2018-06-29

Applicant: AMGEN INC.

Inventor： Da REN ,  Izydor APOSTOL ,  Alexander Julian VEACH

IPC: G01N33/68 ,  B01D15/34

Abstract: The disclosed methods are directed to detecting polypeptide fragments (“clips”) of parental polypeptides. Parental polypeptides and clips are optionally denatured and then fractionated using a matrix. After a first elution (high molecular weight fraction), an additional elution step retrieves a low molecular weight fraction containing clips. If the clips are an appropriate size for the targeted detection method, such as mass spectrometry, then analysis of this fraction proceeds separately from the high molecular weight fraction, or the clips fraction is mixed with the proteolyzed high molecular weight fraction before analysis. However, if the clips are too large for the intended analytical method(s), then the clips are also proteolyzed. The digested high molecular weight and low molecular weight fractions can be analyzed separately or combined. Analysis of combined samples favors clip quantitation because the clips are analyzed together with the remaining counterpart of the parental polypeptide.